水稻突变体pe-1对弱光胁迫的响应机制

doi:10.11983/CBB24039

植物学报 ›› 2024, Vol. 59 ›› Issue (4): 574-584.DOI: 10.11983/CBB24039 cstr: 32102.14.CBB24039

水稻突变体pe-1对弱光胁迫的响应机制

黄佳慧, 杨惠敏, 陈欣雨, 朱超宇, 江亚楠, 胡程翔, 连锦瑾, 芦涛, 路梅, 张维林^*(), 饶玉春^*()

浙江师范大学生命科学学院, 金华 321004

收稿日期:2024-03-11 接受日期:2024-05-07 出版日期:2024-07-10 发布日期:2024-07-10
通讯作者: *E-mail: wlzhangwhu@163.com; 饶玉春, 博士/博士后, 教授, 博士生导师, “双龙学者”特聘教授, 浙江省“高校领军人才培养计划”高层次拔尖人才(2022年), 金华市青年拔尖人才(2023年), 浙江省发明协会理事、植物遗传专业委员会副主任, 中国作物学会会员, 浙江省遗传学会会员。主要从事水稻等禾谷类作物的分子遗传学研究。以第一作者或通讯作者身份在Science Bulletin和New Phytologist等权威期刊上发表论文80余篇,以合作者身份在Nature Plants、Proceedings of the National Academy of Sciences of the United States of America等杂志上发表论文30余篇。主持转基因国家重大专项子课题、国家自然科学基金、浙江省自然科学基金等科研项目。授权国家发明专利25项(第一完成人)。E-mail: ryc@zjnu.cn
基金资助:
浙江省自然科学基金重点项目(LZ23C130003);2024年国家级大学生创新创业训练计划和2024年浙江省大学生科技创新活动计划暨新苗人才计划

Response Mechanism of Rice Mutant pe-1 to Low Light Stress

Jiahui Huang, Huimin Yang, Xinyu Chen, Chaoyu Zhu, Yanan Jiang, Chengxiang Hu, Jinjin Lian, Tao Lu, Mei Lu, Weilin Zhang^*(), Yuchun Rao^*()

College of Life Sciences, Zhejiang Normal University, Jinhua 321004, China

Received:2024-03-11 Accepted:2024-05-07 Online:2024-07-10 Published:2024-07-10
Contact: ^*E-mail: wlzhangwhu@163.com; ryc@zjnu.cn

摘要/Abstract

摘要： 以γ射线诱变籼稻双科早(Oryza sativa subsp. indica cv. ‘Shuangkezao’)获得的早熟鲜绿突变体pe-1为实验材料, 在三叶期和分蘖期进行弱光胁迫, 探讨pe-1与野生型在形态特征、非生物胁迫相关酶活性及其调控基因表达量、叶绿素含量、叶绿体合成与降解及光形态建成相关基因表达对弱光响应的差异。结果表明, 与野生型相比, 弱光胁迫后, pe-1叶片黄化程度显著降低, 株高和叶面积显著增加; 三叶期和分蘖期的叶片中不同叶绿素含量变化不同。此外, pe-1叶绿素含量增加, 且其抗氧化应激反应相关酶过氧化氢酶(CAT)和过氧化物酶(POD)的活性及相关基因的表达量均高于野生型, 表明在弱光胁迫下pe-1活性氧清除能力增强, 适应能力更强。pe-1的光形态建成相关基因表达量高于野生型, 表明弱光处理下pe-1的光接收能力更强。综上, pe-1突变体具有抵御弱光胁迫的潜力, 该结果有助于耐弱光水稻品种的选育。

关键词: 水稻, pe-1, 弱光胁迫, 光形态建成, 叶绿素, 叶绿体合成与降解

Abstract: This study utilized the γ-ray-induced early-maturation, fresh-green mutant line pe-1 from indica rice as an experimental material. At the trilobal stage and the tillering stage, we observed differences in morphological characteristics between pe-1 and wild type. In addition, we measured the activity of antioxidant-related enzymes and their regulatory genes expression, chlorophyll content and chloroplast synthesis and degradation-related gene expression, and photomorphogenesis-related gene expression to detect the differences in the low light response between the pe-1 and wild type. The results showed that pe-1 exhibited less leaf yellowing, taller stature, and larger leaf area compared to wild type post-stress. The changes in chlorophyll content differed between leaves at the trilobal stage and the tillering stage. Additionally, pe-1 resulted in increased chlorophyll content and elevated levels of the stress-responsive enzymes catalase and peroxidase, as well as increased expression of related genes. This indicates enhanced reactive oxygen species sca- venging and stronger adaptability to adverse conditions under low light conditions. Moreover, pe-1 exhibited increased expression levels of genes associated with photomorphogenesis, indicating superior light perception ability under low light intensities. In summary, the pe-1 mutant shows immense potential for survival under low light stress, contributing to the breeding rice with low light tolerance.

Key words: rice, pe-1, low light stress, photomorphogenesis, chlorophyll, chloroplast synthesis and degradation

黄佳慧, 杨惠敏, 陈欣雨, 朱超宇, 江亚楠, 胡程翔, 连锦瑾, 芦涛, 路梅, 张维林, 饶玉春. 水稻突变体pe-1对弱光胁迫的响应机制. 植物学报, 2024, 59(4): 574-584.

Jiahui Huang, Huimin Yang, Xinyu Chen, Chaoyu Zhu, Yanan Jiang, Chengxiang Hu, Jinjin Lian, Tao Lu, Mei Lu, Weilin Zhang, Yuchun Rao. Response Mechanism of Rice Mutant pe-1 to Low Light Stress. Chinese Bulletin of Botany, 2024, 59(4): 574-584.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB24039

https://www.chinbullbotany.com/CN/Y2024/V59/I4/574

图/表 10

表1 PCR反应体系

Table 1 PCR reaction system

Component	Volume (μL)
500 ng∙µL^-1 cDNA	1
10 µmol∙L^-1forward primer	1
10 µmol∙L^-1reverse primer	1
ddH₂O	2
2× SYBR Mix	5

表2 PCR反应程序

Table 2 PCR reaction procedure

Reaction temperature (°C)	Function	Reaction time	Cycle number
95	Predegeneration	5 min	1
95	Denaturation	$ \left.\begin{array}{l} 5 \mathrm{~s} \\ 20 \mathrm{~s} \\ 20 \mathrm{~s} \end{array}\right\}$	39
55	Annealing
72	Extension
72	Final extension	10 min	1
15	Hold	Indefinite	1

表3 qRT-PCR所用引物序列

Table 3 Primer sequences for qRT-PCR

Traits	Primer name	Forward primer (5′-3′)	Reverse primer (5′-3′)
	OsActin	TGGCATCTCAGCACATTCC	TGCACAATGGATGGGTCAGA
Chloroplast synthesis	LOC_Os03g20700	GGGAACTTGGCGTTTCATTA	TCAAATGTCTTGCGTTGCTC
and degradation	LOC_Os03g59640	AAAGAATGGCTCGAAAAGCA	GCCACCACATGGTAGCTTCT
	LOC_Os08g06630	TGGATCAACAAGCATTTCCA	GACAACCCGAATTGTCTGCT
	LOC_Os03g19510	GACCCGTGGAGGATGATATG	GCTGTATCGCTTCCCTTTTG
	LOC_Os03g27770	ACCGACGTCGCCTATGTTTA	TATAGCCACCCCTCCAGTTG
	LOC_Os05g28200	AAGCAGAATGGATGGATTGG	GAGTGACTGAAGCCCCAGAG
	LOC_Os03g45194	TGGTGCAACGAAGAGAAGTG	ACGAGATAATCCGCAACCAC
	LOC_Os06g07210	AAGCAAATCTGCTGGAGGAA	AAAAATATCCGCATGCCAAG
Enzyme activity	LOC_Os02g02400	GGCGTCAACACCTACACCTT	GTCGAACCTCTCCTCCTCCT
	LOC_Os06g51150	GTGAATGCACCAAAATGTGC	CCAGCCTGTTGGAAATTGTT
	LOC_Os03g03910	CCACCACAACAACCACTACG	AGTGCGTCGATCCATCTCTT
	LOC_Os03g57220	CCTCTGGAGCTGAGGATGAG	CCCTGGCTGTTGCATACTCT
	LOC_Os04g53210	GGCTGAATTGGCTGGTTTTA	GTCCGTCCAAGACAGAGAGC
	LOC_Os07g05820	ATTGCGCTTACGGTGGATAC	GATCACTCCCTTCACCAGGA
	LOC_Os01g17260	GGAAGACACCAGCTGAGAGG	CAAGAGACCCAGCCAATGTT
Photomorphogenesis	LOC_Os05g11510	GAGCCACGCGTACTTCTTCT	GGTGGTCGCTCTTGGATG
	LOC_Os02g10860	GCCAAAAAGCTCTCATGGTC	GGAAGGAGAAGGGATCGAAG
	LOC_Os03g51030	AAGAGCTTGCCTTGGAATGA	GGGACAGTTGCTGTTTCCAT
	LOC_Os03g19590	CTCGCCTACTCCGAGAACAC	AGGATGGCGTAGAAGGGTTT
	LOC_Os02g36380	CGTCTACGTCAGAGGCATCA	CTTCCCAGCCTGTTGTCAAT
	LOC_Os04g37920	GATGAAGCAGCTTGTGTGGA	AACAAGGGGGTAACCAGTCC
	LOC_Os06g40080	TCAAGGAACAGGGTCACACA	TGCCCTCCCATTTGTAGAAC

图1 水稻叶片生理表型 (A) 三叶期叶色对比(bar=1.2 cm); (B) 分蘖期叶色对比(bar=2 cm); (C) 三叶期叶片二氨基联苯胺(DAB)染色(bar=1.2 cm); (D) 分蘖期叶片DAB染色(bar=2 cm); (E), (F) 三叶期叶绿体自发荧光(箭头所示) (bars=25 μm); (G), (H) 分蘖期叶绿体自发荧光(bars=100 μm)。WT: 野生型

Figure 1 Physiological phenotype of rice leaves (A) Comparison of leaf color at the trilobal stage (bar=1.2 cm); (B) Comparison of leaf color at the tillering stage (bar=2 cm); (C) Diaminobenzidine (DAB) staining of leaves at the trilobal stage (bar=1.2 cm); (D) DAB staining of leaves at the tillering stage (bar=2 cm); (E), (F) Arrows indicate chloroplast autofluorescence at the trilobal stage (bars=25 μm); (G), (H) Chloroplast autofluorescence at the tillering stage (bars=100 μm). WT: Wild type

图2 弱光胁迫后三叶期水稻株高、叶长和叶宽 ** P<0.01 (Student’s t检验)

Figure 2 Plant height, leaf length and leaf width of rice in the trilobal stage after low light stress ** P<0.01 (Student’s t-test)

图3 弱光胁迫下水稻叶片抗氧化酶系统 (A)-(D) 三叶期; (E)-(H) 分蘖期。* P<0.05; ** P<0.01 (Student’s t检验)

Figure 3 Antioxidant enzymes of rice leaves under low light stress (A)-(D) At the trilobal stage; (E)-(H) At the tillering stage. * P<0.05; ** P<0.01 (Student’s t-test)

图4 水稻酶活性相关基因表达差异 (A) 三叶期; (B) 分蘖期。* P<0.05; ** P<0.01 (Student’s t检验)

Figure 4 Differences in the expression of enzyme activity related genes in rice (A) At the trilobal stage; (B) At the tillering stage. * P<0.05; ** P<0.01 (Student’s t-test)

图5 弱光胁迫下水稻叶绿素含量变化 (A)-(D) 三叶期; (E)-(H) 分蘖期。* P<0.05; ** P<0.01 (Student’s t检验)

Figure 5 Changes in chlorophyll content of rice under low light stress (A)-(D) At the trilobal stage; (E)-(H) At the tillering stage. * P<0.05; ** P<0.01 (Student’s t-test)

图6 水稻叶绿体合成与降解相关基因表达差异 (A) 三叶期; (B) 分蘖期。* P<0.05; ** P<0.01 (Student’s t检验)

Figure 6 Differences in the expression of genes related to chloroplast synthesis and degradation in rice (A) At the trilobal stage; (B) At the tillering stage. * P<0.05; ** P<0.01 (Student’s t-test)

图7 水稻光形态建成相关基因表达差异 (A) 三叶期; (B) 分蘖期。* P<0.05; ** P<0.01 (Student’s t检验)

Figure 7 Differences in the expression of genes related to photomorphogenesis in rice (A) At the trilobal stage; (B) At the tillering stage. * P<0.05; ** P<0.01 (Student’s t-test)

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水稻突变体pe-1对弱光胁迫的响应机制

Response Mechanism of Rice Mutant pe-1 to Low Light Stress

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